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Keywords:

Artemia, Cell Line, Chromosome Mapping, Cloning, Molecular, Cytoplasm, DNA, Gene Expression Regulation, Humans, Melanoma, Multigene Family, Peptide Elongation Factor 1, Peptide Elongation Factors, Phorbol Esters, Sequence Homology, Nucleic Acid, Science & Technology, Life Sciences & Biomedicine, Genetics & Heredity, 0604 Genetics, 1104 Complementary and Alternative Medicine, 1114 Paediatrics and Reproductive Medicine, 3105 Genetics, 3215 Reproductive medicine

Abstract:

One of the genes activated in human melanoma cells by the tumor-promoting phorbol ester is that of the elongation factor 1 alpha. A cDNA clone containing the complete 3'-end untranslated region and the nucleotide sequences coding for 227 carboxyterminal amino acids was isolated. Computer-assisted comparison with known sequences of elongation factors from other species revealed homologies up to 73% and 63% on amino acid and nucleotide sequences, respectively. Northern blot analysis of mRNA from unstimulated and phorbol ester-treated cells showed a 3- to 5-fold increase in cytoplasmic elongation factor 1 alpha mRNA after phorbol ester induction. When compared with the phorbol ester-inducible single-copy gene transcripts coding for the tissue-type plasminogen activator, the cellular mRNA content of elongation factor 1 alpha is 30 times higher. By Southern blot analysis experiments on human genomic DNA, a multi-gene family was found showing polymorphisms in restriction endonuclease fragment lengths (RFLP). Several polymorphisms were studied more extensively in the population on more than 100 DNA samples from normal individuals and in three-generation families. In situ hybridization of the cDNA probe to normal human metaphase chromosomes showed multiple chromosomal localizations of the elongation factor gene(s), with peak hybridization on the chromosomes 1, 2, 4, 5, 6, 7, and 15. The estimate of the gene copy number in humans is more than ten copies per (haploid) genome.